Donald J. Dierschke

Endocrine and ultrasound evaluation of the response to PGF 2α and GnRH given at different stages of the luteal phase in cyclic ewes

To investigate the effects of prostaglandin (PGF 2alpha) plus GnRH at different stages of the luteal phase 13 ewes received PGF 2alpha on Day 9 of the synchronized cycle, followed 36 h later by GnRH. This control regimen resulted in ovulation and normal corpus luteum (CL) function. In the next cycle, the ewes were treated simultaneously with PGF 2alpha and GnRH either on Day 4 (early, n = 7) or Day 9 (late, n = 6). Ovarian activity was monitored daily by ultrasonography, and blood samples were obtained to monitor hormonal patterns. Size of the largest follicle present when GnRH was administered was similar in all groups, but the preceding growth rate was greatest for the early group. In the 36 h after injection of PGF 2alpha, serum progesterone (P4) had declined to basal levels in the control cycles when GnRH was administered, but P4 concentrations were higher in the early group and were highest in the late group when the GnRH was administered with PGF 2alpha. The LH surges induced by GnRH were highest in the control cycles, and were lower in the 2 treated groups. In the early group, 6 of 7 ewes demonstrated ovulation within 48 h of GnRH, resulting in the formation of normal CL. In the late group, ovulation was delayed for about 5 d in 4 of 6 ewes, and subsequent luteal function was normal; no ovulation was detected in the other 2 ewes of this group, but the follicles became luteinized, resulting in a normal P4 profile in one and subnormal in the other. These results suggest that follicles present during the early luteal phase are capable of ovulating and forming fully functional CL in response to exogenous GnRH. In contrast, follicles present during the late luteal phase fail to ovulate in response to GnRH while P4 levels are high, even though the LH stimulus is adequate; however, these follicles persist and subsequently ovulate after P4 levels have decreased. Therefore, the endocrine milieu to which a follicle was exposed may be more important than its size in determining its ability to undergo ovulation and development into a normal CL.

Evaluation of ultrasonography for monitoring follicular growth in rhesus monkeys

The purpose of this study was to determine if the ovaries and uterus of rhesus monkeys could be visualized by ultrasonography and to detect changes associated with follicular growth and ovulation. Animals were examined during 15 menstrual cycles, for an average of nine consecutive days. Ultrasonic recordings were correlated with hormonal parameters (estradiol 17beta, E(2); luteinizing hormone, LH; and progesterone, P) and laparoscopic findings. The uterus and both ovaries were observed in more than 90% of the examinations. A dominant follicle (DF) was identified during all ovulatory cycles, on average 1 d preceding the E(2) peak. The maximal diameter of the DF ranged from 3 to 7 mm. Laparoscopic examinations to determine the site of the DF confirmed ultrasonic findings in 10 of 14 cycles (P < 0.1). There was no significant difference in the size of the dominant and contralateral ovaries; however, more follicles with a diameter of 2 to 7 mm were found on the dominant ovary (P < 0.05). Two animals stimulated with exogenous gonadotropins showed a linear increase in ovarian size for 6 d prior to oocyte recovery (P < 0.05), reflecting an increase in the number of developing follicles. Ultrasonography can be used to identify the DF during spontaneous cycles in rhesus monkeys and to monitor the response of monkeys to exogenous gonadotropins.

Functional Anatomy of the Testicular Vascular Pedicle in the Rhesus Monkey: Evidence for a Local Testosterone Concentrating Mechanism

Summary The detailed anatomy of arteries and veins in the testicular pedicle of the rhesus monkey, with special emphasis on an area of extensive surface contact between these vessels in the spermatic cord, is described. The mean plasma testosterone concentration in blood from the testicular artery was significantly greater than the mean for samples collected simultaneously from the contralateral femoral artery in 11 monkeys. These observations are interpreted as supportive of the hypothesis that a hormone concentrating mechanism involving the local transfer of testosterone between the vessels of the pampiniform plexus and the spermatic artery exists in the male rhesus monkey. The authors gratefully acknowledge the contributions of Dr. P. T. K. Toivola in developing surgical procedures and his suggestions in preparing the manuscript. Photographs were taken by Mr. W. P. Steffenhagen. Expert technical assistance was provided by R. Abrams, G. Scheffler, J. Scheffler, and A. Mitchell.

Role and site of estrogen action in follicular atresia

Subsequent to an initial understanding that estrogen was only stimulatory to folliculogenesis, we have come full circle to the present recognition that many actions of estrogen are inhibitory to follicular function. The development of this interpretation has frequently been associated with the controversial issue o f the likely site o f estrogen action, especially in primates, where much of the evidence has been amassed. The accumulated findings in a variety of species seem to demonstrate clearly that at least part of the atretogenic effect of estrogen is exerted directly on the ovary, apparently by interaction with the nuclear estrogen receptor. Recent observations include identification of messenger RNAs for the estrogen receptor and for creatine kinase in the macaque ovary, and a current focus is to localize messages within specific compartments of the ovary.

Estrogen - Induced release of luteinizing hormone in prepubertal and postpubertal heifers.

This experiment was designed to determine the age at which estradiol-17beta (E(2)) first induces a preovulatory-like surge of luteinizing hormone (LH) in prepubertal heifers. Responses of prepubertal animals 3 to 4 and 5 to 6 months of age were compared with those of postpubertal heifers that received 25 mg prostaglandin F(2)alpha at 0800 hr on day 15 of the estrous cycle. E(2) (500mug) induced surges of LH in 1 5 heifers 3 to 4 months of age, 3 3 heifers 5 to 6 months of age and 5 5 postpubertal heifers. Duration of response and interval between E(2) injection and peak of the response were longer in postpubertal heifers than in those 5 to 6 months old (P<0.10). Peak response and total amount of LH released were greater in animals 5 to 6 months old (P<0.10). Only one prepubertal heifer had elevated concentrations of progesterone following an LH surge. Four of 5 postpubertal heifers receiving E(2) and 3 of 4 postpubertal heifers receiving corn oil had corpora lutea and similar patterns of progesterone concentrations. We conclude that ability to release an LH surge in response to E(2) develops in heifers between 3 and 5 months of age, but that this induced surge does not cause ovulation.

Diminished steroidogenic response of hamster granulosa cells to estrogen in vitro.

SummaryWe have previously demonstrated that estrogen can exert inhibitory or atretogenic effects on the ovaries of both rats and rhesus monkeys in vivo. This study was designed to test whether the hamster (Mesocricetus auratus) is an appropriate model in which to test the effects of estrogens (diethylstilbestrol and estradiol-17β) on steroid accumulation by ovarian granulosa cells in vitro, and whether the effects are similar to those demonstrated for other species in vivo. Immature female hamsters were injected with pregnant mares serum gonadotropin at 28 to 30 days of age. Animals were sacrificed and follicular contents aspirated three days later. Granulosa cells were either left untreated or treated with diethylstilbestrol or estradiol (1×10-7 M) in vitro for 72 h in the presence of androstenedione (1×10-7 M), and in the presence or absence of serum (10%) or human follicle-stimulating hormone (20 ng/ml), and long-term accumulation of estrogen and progesterone was determined. Diethylstilbestrol inhibited accumulation of estrogen regardless of the presence or absence of follicle-stimulating hormone. In contrast, only estradiol plus follicle-stimulating hormone augmented accumulation of progesterone by granulosa cells. These findings that estrogen can be non-stimulatory or inhibitory to function of granulosa cells in vitro parallel those shown in vivo. Our experimental approach may therefore represent an appropriate model for study of the direct effects of estradiol on the function of granulosa cells.

Serum progesterone and corpus luteum function in pregnant pigtailed monkeys (Macaca nemestrina)

Corpus luteum (CL) function and control during pregnancy and early lactation in the pigtailed macaque was investigated. Peripheral concentrations of progesterone (P) on day 10 of pregnancy were 12.98 +/- 2.21 ng/ml and decreased progressively to 7.96 +/- 1.27 ng/ml by day 21 of pregnancy. The concentration of P increased around day 27 of gestation and reached peak levels of 18.48 +/- 2.45 ng/ml on day 37, thereafter gradually decreasing to a nadir at about midgestation. Ten days before parturition P concentrations increased again (P < 0.05). Concentrations of P decreased from 6.62 +/- 1.48 ng/ml on the day of delivery to 2.16 +/- 0.43 ng/ml on day 2 of lactation and remained low thereafter. Ovariectomy on day 35 did not affect the normal course of gestation or the patterns of P secretion during pregnancy. However, in these ovariectomized animals, in spite of suckling, P was not detectable after parturition. In intact monkeys, serum concentrations of P in the utero-ovarian vein at days 80 and 159 of pregnancy were higher relative to the uterine vein. Incubation studies utilizing 3H-cholesterol as a substrate revealed that the CL were capable of synthesizing P on days 35 and 159 of gestation. Histologically, the CL contained active luteal cells at late pregnancy. Low serum concentrations of chorionic gonadotropin were detected on day 10 of gestation; concentrations of this hormone reached high levels between days 18 and 24 and the titers were nondetectable after day 40 of pregnancy. Luteinizing hormone was present in constant amounts in the circulation during pregnancy and lactation. These data suggest that the CL of pregnancy in the pigtailed monkey is functional or capable of functioning during various stages of pregnancy. However, the fetoplacental unit is the primary source of P during the latter 4.5 months of gestation. As in other primates, a functional CL is not required for maintenance of pregnancy after implantation nor for lactation. Thus, the physiological significance of CL function during pregnancy is unclear.

Influences of month of birth and age on patterns of luteinizing hormone secretion in prepubertal heifers.

An experiment was done to determine if month of birth and age influenced patterns of luteinizing hormone (LH) secretion in prepubertal heifers. Mean concentrations of LH increased linearly (P<.05) in March-born heifers between one and seven months of age. This was partially due to an increase in number of LH pulses. The prepubertal pattern of LH concentrations was quadratic (P<.05) for heifers born in September because concentrations were slightly higher (P=.15) than those in March-born heifers at one month of age. There were no differences between groups during the remainder of the prepubertal period (3 to 7 months). There was a tendency (P=.18) for September-born animals to reach puberty at younger ages than those born in March. September-born heifers also had greater (P=.06) average daily gains, but body weights at puberty were similar for the two groups. These results show that season of birth influenced LH concentrations at one month of age, but did not significantly affect the increase between three and seven months of age.

FSH-induced aromatase activity in hamster granulosa cells: effect of estradiol-17β in vitro

SummaryWe have shown previously that estradiol-17β (E2) reduces number of ovulations in cyclic rats, induces atresia of the dominant preovulatory follicle in monkeys, and that the initial effects of this treatment include reduced viability and estrogen accumulation in vitro by aspirated granulosa cells (GC) from monkeys and hamsters. The present experiment was designed to determine whether the reduction in estrogen accumulation can be ascribed to a direct action of E2 on the aromatization of androgen to estrogen in vitro. Female hamsters were injected with 30 I.U. pregnant-mare serum gonadotropin i.p. and sacrificed 3 days later. GC were aspirated from the largest follicles and incubated for 48 h (“initial incubation” period) in the presence of human pituitary follicle-stimulating hormone (hFSH, 100 ng/ml). Following initial incubation, GC were further incubated for up to 24 h (“secondary incubation” period). During this subsequent incubation, medium was supplemented with 100nM 3H-1β-androstenedione (3H-A4). Initial incubation with E2 at doses of 10 ng/ml, 100 ng/ml and 1 μm E2/ml induced variability in GC response, and a maximal depression of ∼70%. The inhibition by E2 of hamster GC function in vitro parallels that shown in vivo for both hamsters and monkeys, but contrasts with that shown for rats. Thus, hamsters may represent an appropriate model in which to study the atretogenic effects of E2 directly on antral follicle development.

Anterior hypothalamic lesions and pubertal development in female rhesus monkeys

Electrolytic lesions were made in the anterior hypothalamus of 8 prepubertal female rhesus monkeys, aged 1.1-1.7 years. Six unoperated females served as controls. No effects were found of the lesions upon age and body weight at menarche or at first ovulation, as estimated by blood levels of progesterone and laparoscopic observations. From these findings it appears that the neural control of puberty in the female rhesus may not be exerted through the anterior hypothalamus, in contrast with the rat and ferret. Further, an attempt was made to identify biometric correlates of hormonal changes during puberty. Firstly, the well known dip in growth rate, about 0.4 years before menarche, was observed. Secondly, there was a marked spurt in growth of the nipples starting at 0.2 years before menarche. The close temporal association between accelerated nipple growth and menarche suggests that both of these developmental characteristics result from changes in (presumably ovarian) steroid hormone secretion.